Cancers Of Upper Aerodigestive Tract Research Articles

Abstract 3861: A genome-wide association study of upper aerodigestive tract cancers identifies 4q21, 4q23, and 12q24 as susceptibility loci

Abstract To identify risk variants for upper aero-digestive tract (UADT) cancers, we performed a genome-wide association study of 2,091 UADT cancer cases and 8,334 controls followed by replication of 20 variants in an additional 6,545 UADT cancer cases and 7,892 controls. Five common variants presented evidence for significant association in the replication series (one sided p ≤ 0.005 - preplication) and combined analysis (p ≤ 5×10−7 - pcombined). Three variants were in the alcohol dehydrogenase (ADH) gene cluster on 4q23 (rs1573496-ADH7, preplication=7×10−10 /pcombined=5×10−17 rs1229984-ADH1B, preplication=3×10−10 pcombined=5×10−21 and rs698-ADH1C, preplication=0.003 pcombined=3×10−7). The fourth variant (rs4767364, preplication=4×10−4/pcombined=7×10−8) was located in an extended linkage disequilibrium region at 12q24 that contains the aldehyde dehydrogenase 2 (ALDH2) gene and appears to have effects similar, albeit more modest, to that observed with heterozygote ALDH2 null allele (rs671) common in Asian populations. The fifth variant (rs1494961, preplication=9×10−6 pcombined=2×10−8) was located near DNA repair related genes HEL308 and FAM175A (or Abraxas) at 4q21. This 4q21 variant was also associated with lung cancer risk in a series of 5,652 lung cancer cases and 9,338 controls (rs1494961, p=3×10−4). These results further highlight the importance of the genes involved in the metabolism of alcohol in UADT cancer susceptibility, and also implicate 4q21 in susceptibility to both UADT and lung cancers. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3861.

Abstract 3865: A post genome-wide association study: Gene and environment interaction in the risk of head and neck carcinoma

Abstract A consortium of case-control studies of upper aerodigestive tract (UADT) cancers recently conducted a genome-wide association study (GWAS) and identified 20 candidate SNPs demonstrating significant independent risk for disease. Although this collaborative effort provided the sample size necessary to identify these candidates, it was not possible to investigate the interaction of these genes with environmental and demographic factors due to differences in the various participating studies’ designs, data collection strategies, and diseases examined. Thus, our study sought to investigate the potential low penetrance gene-gene and gene-environment interaction in head and neck squamous cell carcinoma (HNSCC) risk, we examined the significant variants identified from the initial GWAS and conducted a post-GWAS analysis of gene-environment and higher order gene-gene-environment interactions in our population-based case-control study of HNSCC. Cases were patients with incident HNSCC identified from nine medical facilities in Greater Boston, MA between December 1999 and December 2003. Controls were identified from Massachusetts town books using random selection and frequency-matched to cases on age (± 3 years), gender, and town of residence. There were twenty variants selected from initial GWAS for UADT cancers. These included 11 variants with p-values <10−5, 6 non-synonymous SNPs with p-values <10−4 and 2 variants with p-values <5×10−7 in restricting to UADT cancer subsite, or heavy drinkers/smokers. Finally, a non-synonmous SNP (rs1229984) was included. Taqman (Applied Biosystems) genotyping assays were designed and utilized to genotype the 575 cases and 676 controls. A restricted cubic spline model suggested both ADH1B and HEL308 modified the association between smoking pack-years and HNSCC. Subanalysis suggested the departure from multiplicative interaction between ADH1B and smoking was significant consistently across tumor sites (oral, pharynx and larynx). However, the departure from multiplicative interaction between HEL308 and smoking was only detected in oral cavity and pharyngeal sites. Classification and regression tree (CART) analysis demonstrated the higher order interactions between smoking status, ADH1B, FLJ13089 and FLJ35784 in HNSCC risk. Compared with ever smokers carrying ADH1B T/C+T/T genotypes, smokers carrying ADH1B C/C genotype and FLJ13089 A/G+A/A genotypes had a highest risk of HNSCC (OR=1.84; 95%CI: 0.98-3.47). Our study showed both ADH1B and HEL308 modified the association between smoking and HNSCC. Higher order interactions between smoking status, ADH1B, FLJ13089 and FLJ35784 in HNSCC risk were also observed. Together these results confirm and expand upon the associations observed in the initial GWAS and suggest that the risk associated with these variants may be specifically important amongst specific exposure groups. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3865.

Abstract 2818: Vitamin or mineral supplement intake and the risk of head and neck cancer: Pooled analysis in the INHANCE consortium

Abstract Background. Head and neck cancer (HNC) is a significant cause of morbidity and mortality, with over half a million cases worldwide each year. While high fruit and vegetable intake is thought to be protective against HNC, it is unclear whether vitamin and mineral supplement intake is associated with decreased risk of these malignancies. Previous case-control studies in the US have suggested vitamin E supplement intake is inversely associated with oral cavity and pharyngeal cancers. In a large randomized controlled trial, supplementation with alpha-tocopheryl acetate and/or beta-carotene was not associated with upper aerodigestive tract cancer incidence; however an inverse association was suggested for early stage laryngeal cancers. Methods. We analyzed individual-level pooled data from 12 case-control studies (7,002 HNC cases and 8,383 controls) participating in the International Head and Neck Cancer Epidemiology (INHANCE) consortium. There were a total of 2,028 oral cavity cancer cases, 2,465 pharyngeal cancer cases, and 874 laryngeal cancer cases. Odds ratios (OR) and 95% confidence intervals (CIs) for ever use of any vitamins, multivitamins, vitamin A, vitamin C, vitamin E, and calcium, beta carotene, iron, selenium, and zinc supplement were assessed. We further examined frequency, duration and cumulative exposure of each vitamin or mineral when possible. All ORs were adjusted for age, sex, race/ethnicity, study center, education level, pack-years of smoking, frequency of alcohol drinking and fruit/vegetable intake. Results. A decreased risk of HNC was observed with ever use of vitamin C (OR=0.76, 95%CI=0.59-0.96) and with ever use of calcium supplements (OR=0.64, 95%CI=0.42-0.97). There did not appear to be any associations between HNC risk and ever use of any vitamins (OR=0.87, 95%CI=0.71-1.06), multiple vitamins (OR=0.99, 95%CI=0.82-1.19), vitamin A (OR=1.04, 95%CI=0.54-1.98), vitamin E (OR=0.71, 95%CI=0.45-1.11), beta-carotene (OR=1.35, 95%CI=0.65-2.81), iron (OR=0.79, 95%CI=0.54-1.16), selenium (OR=1.21, 95%CI=0.35-4.22) or zinc (OR=0.88, 95%CI=0.34-2.26). Dose-response trends were suggested for years of use of vitamin C (p for trend=0.03) and calcium (p<0.01) with decreasing HNC risk, but not for other vitamin or mineral supplement use. Increased cumulative calcium intake (in tablets) was also associated with decreasing HNC risk (p for trend<0.01). Differences by cancer subsite (oral cavity, pharynx, larynx) were not observed. Conclusions. This is the first time that inverse associations were observed for HNC risk with both vitamin C use and with calcium use in an extremely large sample size. The next steps for our pooled analyses will include adjustment on micronutrients from dietary intake and stratification on HPV infection status where possible. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2818.

Abstract 4174: Cxcl5 and Cxcl2 overexpression in esophageal carcinogenesis is associated with rapid tumor formation in zinc-deficient rats

Abstract Objectives: Zinc-deficiency (ZD) is implicated in the pathogenesis of human esophageal cancer. It induces cell proliferation, modulates genetic expression, and enhances carcinogenesis. Zinc-replenishment (ZR) reverses proliferation and inhibits carcinogenesis. Recently, we showed that a ZD diet induces overexpression of the proinflammation mediators S100a8/a9 in hyperplastic rat esophagus, providing evidence that zinc regulates an inflammatory pathway in early esophageal carcinogenesis (Gastroenterology, 136: 953-966, 2009). Here we determined the biological processes affected by zinc nutrition during rapid N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumor development and its prevention. Methods: We evaluated gene expression profiles of esophageal mucosa from zinc-modulated rats at 5 weeks after an NMBA dose and at 15 weeks (endpoint) with 3 NMBA doses, using Affymetrix Rat Genome GeneChip. Here, ZD rats documented a significantly higher tumorigenic outcome than control zinc-sufficient (ZS) rats (tumor incidence, 100% vs 16.6%; multiplicity, 11 ± 3.8 vs 0.5 ± 0.3, P<0.001). Replenishing zinc after the first NMBA dose led to a low tumorigenic outcome in ZR vs ZD rats (incidence, 28.9% vs 100%; multiplicity, 0.6 ± 0.4 vs 11 ± 3.8, P<0.001). Results: The ZD esophagi have a distinct expression signature versus ZS esophagi at both time points. At 5 weeks, in addition to S100a8/a9 overexpression (up 2.5-fold), the dysplastic ZD esophagi showed up-regulation of several cancer-related inflammation genes, including CXC chemokines Cxcl5 and Cxcl2 (up >25-fold); Ptgs2 (up 26-fold); and Il17f (up 2.7 fold). At endpoint, the tumor-bearing ZD esophagus displayed a neoplastic gene signature, with increased up-regulation of Cxcl5 and Cxcl2 (up 84- and 38-fold), Cxcl3 (up 9-fold), Cxcl1 (up 7-fold), Ptgs2 (up 41-fold), Il1b (up 12-fold), Il17f (up 7.5-fold), and S100a8/a9 (up 2.1-fold). Replenishing zinc led to a global reversal of abnormal gene expression, resulting in an expression profile similar to that of ZS esophagi. In parallel to mRNA expression, CXCL5, CXCL2, COX-2, and S100A8/A9 protein expression was similarly modulated by ZD and ZR, as shown by ELISA, immunoblotting, and immunohistochemistry assays. Ingenuity Pathway Analysis predicted an Il1b-centric network, with direct connections to many up-regulated genes. Conclusions: ZD fuels inflammation and drives carcinogenesis by inducing overexpression of several cancer-related inflammation genes, including Cxcl5 and Cxcl2. ZR modulates inflammatory responses and inhibits tumor growth. The data provide a new understanding of the molecular role of zinc in esophageal carcinogenesis/prevention and suggest that zinc supplementation should be more thoroughly explored in human prevention clinical trials for upper aerodigestive tract cancer. Supported by NIH grant CA118560. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4174.

Alcohol drinking, cigarette smoking, and the development of squamous cell carcinoma of the esophagus: epidemiology, clinical findings, and prevention

Both cigarette smoking and alcohol drinking are well-established risk factors for esophageal squamous cell carcinoma (ESCC), and the relationship of dose to cancer risk has already been described. Furthermore, the synergistic effect of these two factors has been reported. Our case-control study revealed the odds ratio of ESCC to be 50.1 for those who were both heavy smokers and heavy drinkers in comparison to people who neither drank nor smoked. In patients with ESCC, head and neck cancers as well as dysplastic lesions are frequently observed. Heavy smoking and heavy drinking are closely related to such multicentric carcinogenesis events in the upper aerodigestive tract (UADT), including the esophagus and head andneck region. Polymorphisms in acetaldehyde dehydrogenase 2 (ALDH2) are reported to be a key event in deciding individual susceptibility to UADT cancer. Patients with inactive ALDH2, in whom facial flushing is usually observed after the drinking of alcohol, are at high risk for ESCC as well as multiple UADT cancers. For the early detection of the disease, effective follow up using endoscopy with Lugol staining or narrow band imaging endoscopy is strongly recommended for high-risk populations, such as smokers, heavy drinkers, people with experience of flushing after the drinking of alcohol, and patients with UADT cancer.

Life course social mobility and risk of upper aerodigestive tract cancer in men

The aim of this study was to explore associations between social mobility and tumours of the upper aero-digestive tract (UADT), focussing on life-course transitions in social prestige (SP) based on occupational history. 1,796 cases diagnosed between 1993 and 2005 in ten European countries were compared with 1585 controls. SP was classified by the Standard International Occupational Prestige Scale (SIOPS) based on job histories. SIOPS was categorised in high (H), medium (M) and low (L). Time weighted average achieved and transitions between SP with nine trajectories: H --> H, H --> M, H --> L, M --> H, M --> M, M --> L, L --> H, L --> M and L --> L were analysed. Odds ratios (ORs) and 95%-confidence intervals [95%-CIs] were estimated with logistic regression models including age, consumption of fruits/vegetables, study centre, smoking and alcohol consumption. The adjusted OR for the lowest versus the highest of three categories (time weighted average of SP) was 1.28 [1.04-1.56]. The distance of SP widened between cases and controls during working life. The downward trajectory H --> L gave an OR of 1.71 [0.75-3.87] as compared to H --> H. Subjects with M --> M and L --> L trajectories ORs were also elevated relative to subjects with H --> H trajectories. The association between SP and UADT is not fully explained by confounding factors. Downward social trajectory during the life course may be an independent risk factor for UADT cancers.

Impact of smoking on lung cancer risk is stronger in those with the homozygous aldehyde dehydrogenase 2 null allele in a Japanese population

The main lifestyle contributor to acetaldehyde exposure is the drinking of alcoholic beverages, but tobacco smoke also makes some contribution. Although acetaldehyde is associated with upper aerodigestive tract cancer risk, in accordance with genetically determined acetaldehyde metabolism, it is unclear whether lung cancer, a representative smoking-related cancer, is associated with acetaldehyde or genes impacting its metabolism. We conducted a case-control study to examine possible interaction between smoking and aldehyde dehydrogenase 2 (ALDH2) Glu504Lys polymorphism (rs671) on the risk of lung cancer in Japanese. Subjects were 718 lung cancer cases and 1416 non-cancer controls enrolled in the Hospital-based Epidemiologic Research Program at Aichi Cancer Center. Lifestyle factors, including smoking, were determined by self-administered questionnaire. We applied pack-years (PY; categorized into five levels: never, <15, <30, <45 and > or =45) as a marker of cumulative exposure to smoking. The impact of smoking, ALDH2 genotype, and their interaction on lung cancer risk were assessed by odds ratio (OR) and 95% confidence interval adjusted for potential confounders. Adjusted ORs for PY <15, <30, <45 and > or =45 relative to never smokers among those with Glu/Glu or Glu/Lys were 1.39, 1.80, 3.44 and 6.25, respectively (P-trend = 1.4 x 10(-30)). In contrast, ORs among Lys/Lys were 1.01, 10.2, 11.4 and 23.2, respectively (P-trend = 2.6 x 10(-7)). Interaction between ALDH2 genotype (Glu/Glu + Glu/Lys versus Lys/Lys) and cumulative smoking dose was statistically significant (P = 0.036) and was consistently observed in the analysis among never-drinkers (interaction P = 0.041). These results suggest that ALDH2 Lys/Lys, a null enzyme activity genotype, modifies the impact of smoking on the risk of lung cancer.

Alcohol, Tobacco and Genetic Susceptibility in Relation to Cancers of the Upper Aerodigestive Tract in Northern Italy

Each year in Italy there are approximately 14,000 new cases and 7,000 deaths from cancer of the upper aerodigestive tract, which includes malignant tumors originating from the oral cavity, pharynx, larynx and esophagus. Established etiological factors include tobacco consumption and heavy alcohol drinking. The study of single nucleotide polymorphisms in upper aerodigestive tract cancer etiology may help to identify high-risk subgroups and to better understand the pathways leading to the development of these cancers. Italian results on about 500 cases and 500 controls from a large case-control study (ARCAGE) conducted in 10 European countries are presented with the major objectives of updating results on the effects of alcohol and tobacco consumptions in northern Italy, investigating the role of genetic variation with regard to the metabolism of alcohol and carcinogens from tobacco smoke, and evaluating possible interactions of these single nucleotide polymorphisms with these carcinogens. The present study confirmed the importance of tobacco smoking and alcohol drinking as the main risk factors for upper aerodigestive tract cancers, indicating that about 68% of cancers among populations in northern Italy can be attributed to the combination of these risk factors. Significant associations between metabolizing phase I genes (CYP1A1 and CYP2A6), phase II genes (GSTA2) and upper aerodigestive tract cancers were found. A polymorphism of ADH1C has been associated with an increased risk of upper aerodigestive tract cancers, suggesting that the less rapid alcohol metabolizers are more susceptible to upper aerodigestive tract cancer risk. Our results suggest that the ADH1C allele modifies the carcinogenic dose response for alcohol in the upper aerodigestive tract, giving rise to a gene-environment interaction. The role of genes as possible modifiers of life-style risks seems the most reliable.

Alcohol and Aldehyde Dehydrogenase Polymorphisms and a New Strategy for Prevention and Screening for Cancer in the Upper Aerodigestive Tract in East Asians

The ethanol in alcoholic beverages and the acetaldehyde associated with alcohol consumption are Group 1 human carcinogens (WHO, International Agency for Research on Cancer). The combination of alcohol consumption, tobacco smoking, the inactive heterozygous aldehyde dehydrogenase-2 genotype (ALDH2*1/*2) and the less-active homozygous alcohol dehydrogenase-1B genotype (ADH1B*1/*1) increases the risk of squamous cell carcinoma (SCC) in the upper aerodigestive tract (UADT) in a multiplicative fashion in East Asians. In addition to being exposed to locally high levels of ethanol, the UADT is exposed to a very high concentration of acetaldehyde from a variety of sources, including that as an ingredient of alcoholic beverages per se and that found in tobacco smoke; acetaldehyde is also produced by salivary microorganisms and mucosal enzymes and is present as blood acetaldehyde. The inefficient degradation of acetaldehyde by weakly expressed ALDH2 in the UADT may be cri! tical to the local accumulation of acetaldehyde, especially in ALDH2*1/*2 carriers. ADH1B*1/*1 carriers tend to experience less intense alcohol flushing and are highly susceptible to heavy drinking and alcoholism. Heavy drinking by persons with the less-active ADH1B*1/*1 leads to longer exposure of the UADT to salivary ethanol and acetaldehyde. The ALDH2*1/*2 genotype is a very strong predictor of synchronous and metachronous multiple SCCs in the UADT. High red cell mean corpuscular volume (MCV), esophageal dysplasia, and melanosis in the UADT, all of which are frequently found in ALDH2*1/*2 drinkers, are useful for identifying high-risk individuals. We invented a simple flushing questionnaire that enables prediction of the ALDH2 phenotype. New health appraisal models that include ALDH2 genotype, the simple flushing questionnaire, or MCV are powerful tools for devising a new strategy for prevention and screening for UADT cancer in East Asians.

Abstract A207: Whole transcriptome profiling in upper aerodigestive tract cancers

Abstract Background: RNA-seq is a powerful technology that allows to obtain sequence and expression information simultaneously. We applied this technology to 4 upper aerodigestive tract cancers. Methods: RNA from four upper aerodigestive tract tumor specimens was extracted and sequencing libraries constructed. Samples were analyzed using an Illumina Genome Analyzer with a paired end module (36 or 54 base read length). Raw data was processed with a proprietary data pipeline. Potential mutations were filtered using both in house and external bioinformatic solutions, involving previously established databases of mutations (COSMIC) and SNPs (dbSNP), and removal of ancestral alleles identified from multiple sequence alignments. These SNPs were then parsed via in house scripts to determine whether the SNPs were present in coding regions, 3′UTR, 5′UTR, or in splice acceptor/donor sites. The coding SNPs were further parsed to determine which SNPs result in non-synonymous changes. RNA-Seq expression data was analyzed using Partek Genomics Suite and compared to microarray based expression data. Pathway analysis was performed using GeneGO Metacore and DAVID. Results: 500Mb to 2GB of data were obtained per sample. Between 700 and 2000 nsSNPs were identified, as well as a large number of alteration in the 3′ and 5′ untranslated regions. Genetic alterations in several commonly mutated genes were identified including TP53, ErbB2, H-RAS, and Cyclin D1. SNPs were enriched in pathways commonly involved in upper aerodigestive tract tumors including cell cycle control, cytoskeleton, and receptor tyrosine kinases. Furthermore we screened for viral sequences identifying one sample that was HPV16 positive. RNA-seq based and microarray based expression data generally correlated, but significant differences were also observed, that were at least partly due to the limited number of RNA-seq reads. Conclusion: Cancer transcriptome sequencing is a promising and cost effective approach for identifying mutations and obtaining expression analysis simultaneously. Transcriptome sequencing holds promise as a readily available platform for assessing potential treatment targets in a specific tumor. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):A207.

Gene–environment interactions associated with CYP1A1 MspI and GST polymorphisms and the risk of upper aerodigestive tract cancers in an Indian population

Genetic risk to tobacco related cancers are associated with polymorphisms in CYP1A1 and GST, which are involved in the metabolic activation and detoxification of carcinogens. The genetic variations in these drug-metabolizing enzymes may alter the susceptibility to UADT cancers triggered by environmental exposures. The hospital-based case-control study evaluated the impact of combined CYP1A1 MspI and GST (M1 & T1) polymorphisms among the individuals exposed to environmental risk factors as modulators in the risk of UADT cancers in Tamilians, a population of south India. The unrelated histopathologically confirmed 408 cases and 220 population-based controls matched by age and gender were genotyped for CYP1A1 MspI, GSTM1 and GSTT1 polymorphisms using PCR based methods. To investigate the potential gene-environment interactions, analyses were carried out stratifying by smoking and tobacco chewing status using SPSS software. The combination of genes and environment interactions by stratified analyses revealed significant interactions among the habitual tobacco smokers (CYP1A1 MspI & GSTM1 null: OR 14.06; 95% CI 3.90-50.68, CYP1A1 MspI & GSTT1 null: OR 33.28; 95% CI 4.24-261.19) and tobacco chewers (CYP1A1 MspI & GSTM1 null: OR 20.51; 95% CI 6.77-62.13, CYP1A1 MspI & GSTT1 null: OR 79.35; 95% CI 10.40-605.55) on the multiplicative scale. Our findings have indicated that the individuals polymorphic for CYP1A1 MspI either with GSTM1 null or with GSTT1 null genotypes revealed an increased risk for UADT cancers than that ascribed to a single susceptible gene among the tobacco users in the population [single gene risk among smokers and chewers, respectively, for CYP1A1 MspI (OR 6.43; 95% CI 3.69-11.21); (OR 10.24; 95% CI 5.95-17.60), GSTM1*0 (OR 3.77; 95% CI 1.94-7.37); (OR 7.97 95% CI 4.10-15.76) and GSTT1*0 (OR 6.95 95% CI 2.88-16.77); (OR 25.83 95% CI 7.78-85.76).

Active and Involuntary Tobacco Smoking and Upper Aerodigestive Tract Cancer Risks in a Multicenter Case-Control Study

Several important issues for the established association between tobacco smoking and upper aerodigestive tract (UADT) cancer risks include the associations with smoking by cancer subsite, by type of tobacco, and among never alcohol drinkers and the associations with involuntary smoking among nonsmokers. Our aim was to examine these specific issues in a large-scale case-control study in Europe. Analysis was done on 2,103 UADT squamous cell carcinoma cases and 2,221 controls in the Alcohol-Related Cancers and Genetic Susceptibility in Europe project, a multicenter case-control study in 10 European countries. Unconditional logistic regression was done to obtain odds ratios (OR) and 95% confidence intervals (95% CI). Compared with never tobacco smoking, current smoking was associated with UADT cancer risks (OR, 6.72; 95% CI, 5.45-8.30 for overall; OR, 5.83; 95% CI, 4.50-7.54 for oral cavity and oropharynx; OR, 12.19; 95% CI, 8.29-17.92 for hypopharynx and larynx; and OR, 4.17; 95% CI, 2.45-7.10 for esophagus). Among never drinkers, dose-response relationships with tobacco smoking pack-years were observed for hypopharyngeal and laryngeal cancers (P(trend) = 0.010) but not for oral cavity and oropharyngeal cancers (P(trend) = 0.282). Among never smokers, ever exposure to involuntary smoking was associated with an increased risk of UADT cancers (OR, 1.60; 95% CI, 1.04-2.46). Our results corroborate that tobacco smoking may play a stronger role in the development of hypopharyngeal and laryngeal cancers than that of oral cavity and oropharyngeal cancers among never drinkers and that involuntary smoking is an important risk factor for UADT cancers. Public health interventions to reduce involuntary smoking exposure could help reduce UADT cancer incidence.

Impact of Multiple Alcohol Dehydrogenase Gene Polymorphisms on Risk of Upper Aerodigestive Tract Cancers in a Japanese Population

Alcohol intake is positively associated with the risk of upper aerodigestive tract (UAT) cancer. The genes that encode alcohol-metabolizing enzymes, primarily alcohol dehydrogenases (ADH) and aldehyde dehydrogenases (ALDH), are polymorphic. In Caucasians, significant associations between polymorphisms in ADH1B (rs1229984) and ADH1C (rs698 and rs1693482), and UAT cancer have been observed, despite strong linkage disequilibrium among them. Moreover, UAT cancer was significantly associated with rs1573496 in ADH7, and not with rs1984362 in ADH4. However, little evidence is available concerning ADH4 or ADH7 polymorphisms in Asian populations. We conducted a matched case-control study to clarify the role of ADH polymorphisms in a Japanese population. Cases and controls were 585 patients with UAT cancer and 1,170 noncancer outpatients. Genotyping for ADHs and ALDH2 was done using TaqMan assays. Associations between polymorphisms and UAT cancer were assessed by odds ratios and 95% confidence intervals using conditional logistic regression models that adjusted for age, sex, smoking, drinking, and ALDH2. Adjusted odds ratios were significant for rs4148887 and rs3805322 in ADH4, rs1229984 in ADH1B, rs698 and rs1693482 in ADH1C, and rs284787, rs1154460, and rs3737482 in ADH7. We also observed that ADH7 rs3737482 and ADH4 rs4148887 had independently and statistically significant effects on UAT cancer. The magnitude of effect of these ADH polymorphisms was greater in subjects who were heavy drinkers, heavy smokers, and had esophageal cancer. These findings show that multiple ADH gene polymorphisms were associated with UAT cancer in this Japanese population. Further studies in various ethnicities are required.

Socioeconomic factors associated with risk of upper aerodigestive tract cancer in Europe

Introduction In the European Union, there are 180,000 new cases of upper aerodigestive tract (UADT) cancer cases per year – more than half of whom will die of the disease. Socioeconomic inequalities in UADT cancer incidence are recognised across Europe. We aimed to assess the components of socioeconomic risk both independently and through their influence on the known behavioural risk factors of smoking, alcohol consumption and diet. Patients and methods A multicentre case–control study with 2198 cases of UADT cancer and 2141 controls from hospital and population sources was undertaken involving 14 centres from 10 countries. Personal interviews collected information on demographics, lifetime occupation history, smoking, alcohol consumption and diet. Socioeconomic status was measured by education, occupational social class and unemployment. Odds ratios (ORs) and 95% confidence intervals (CIs) were computed using unconditional logistic regression. Results When controlling for age, sex and centre significantly increased risks for UADT cancer were observed for those with low versus high educational attainment OR = 1.98 (95% CI 1.67, 2.36). Similarly, for occupational socioeconomic indicators – comparing the lowest versus highest International Socio-Economic Index (ISEI) quartile for the longest occupation gave OR = 1.60 (1.28, 2.00); and for unemployment OR = 1.64 (1.24, 2.17). Statistical significance remained for low education when adjusting for smoking, alcohol and diet behaviours OR = 1.29 (1.06, 1.57) in the multivariate analysis. Inequalities were observed only among men but not among women and were greater among those in the British Isles and Eastern European countries than in Southern and Central/Northern European countries. Associations were broadly consistent for subsite and source of controls (hospital and community). Conclusion Socioeconomic inequalities for UADT cancers are only observed among men and are not totally explained by smoking, alcohol drinking and diet.

Association between a 15q25 gene variant, smoking quantity and tobacco-related cancers among 17 000 individuals

Genetic variants in 15q25 have been identified as potential risk markers for lung cancer (LC), but controversy exists as to whether this is a direct association, or whether the 15q variant is simply a proxy for increased exposure to tobacco carcinogens. We performed a detailed analysis of one 15q single nucleotide polymorphism (SNP) (rs16969968) with smoking behaviour and cancer risk in a total of 17 300 subjects from five LC studies and four upper aerodigestive tract (UADT) cancer studies. Subjects with one minor allele smoked on average 0.3 cigarettes per day (CPD) more, whereas subjects with the homozygous minor AA genotype smoked on average 1.2 CPD more than subjects with a GG genotype (P < 0.001). The variant was associated with heavy smoking (>20 CPD) [odds ratio (OR) = 1.13, 95% confidence interval (CI) 0.96-1.34, P = 0.13 for heterozygotes and 1.81, 95% CI 1.39-2.35 for homozygotes, P < 0.0001]. The strong association between the variant and LC risk (OR = 1.30, 95% CI 1.23-1.38, P = 1 x 10(-18)), was virtually unchanged after adjusting for this smoking association (smoking adjusted OR = 1.27, 95% CI 1.19-1.35, P = 5 x 10(-13)). Furthermore, we found an association between the variant allele and an earlier age of LC onset (P = 0.02). The association was also noted in UADT cancers (OR = 1.08, 95% CI 1.01-1.15, P = 0.02). Genome wide association (GWA) analysis of over 300 000 SNPs on 11 219 subjects did not identify any additional variants related to smoking behaviour. This study confirms the strong association between 15q gene variants and LC and shows an independent association with smoking quantity, as well as an association with UADT cancers.

Diet and cancer in Mediterranean countries: carbohydrates and fats

Several aspects of the diet characteristic of the Mediterranean countries are considered favourable not only on cardiovascular disease, but also on cancer risk. We considered some aspects of the Mediterranean diet (including, in particular, the consumption of olive oil and carbohydrates) on cancer risk. Data were derived from a series of case-control studies, conducted in Italy since the early 1990s, on over 10,000 cases of thirteen cancer sites and over 17,000 controls. Olive oil, and other mono- and unsaturated fats, appear to be favourable indicators of breast, ovarian, colorectal, but mostly of upper aero-digestive tract cancers. Whole grain foods are also related to reduced risk of upper aero-digestive tract and various other cancers. In contrast, refined grain intake and, consequently, glycaemic index and glycaemic load were associated to increased risk for several cancer sites. Fish, and hence a diet rich in n-3 polyunsaturated fatty acids, tended to be another favourable diet indicator, while frequent red meat intake was directly related to some common neoplasms. An a priori defined Mediterranean diet score was inversely related to upper digestive and respiratory tract cancers. These data provide additional evidence that major characteristics of the Mediterranean diet favourably affect cancer risk.

8541 Chemoprevention of phytochemicals for head and neck cancers

Cancers of the upper aerodigestive tract (UADT) are mostly carcinomas of mucosal epithelium or secretory gland origin. The majority type is squamous cell carcinoma (SCC), which in USA comprised 95% of UADT cancers registered in the recent SEER registry. Pattern of UADT cancers in Asia has not been known much, although some difference is expected, considering much use of tobacco and alcohol, prevalence of Epstein–Barr virus and different dietary habit. Cases registered in the Surgical Pathology File of Department of Pathology, Korea Cancer Center Hospital, were retrieved from 1988 through 1998. Tumors of the skin, eyelids, eyeball, teeth, ear and brain, benign tumors and repetitive biopsies from the same patients were excluded, and 2842 cases were subjected to pathologic review and necessary reclassification. The tumors were analyzed according to the locations and histologic types. The larynx was the most common site (26%), followed by oral cavity (25%) and oropharynx (13%). Carcinoma comprised 87.8% of total cases, followed by malignant lymphoma (9.0%), sarcoma (1.5%) and malignant melanoma (1.4%). SCC and variants accounted for 76.5%. Sinonasal cancers provided the most diverse histologic types, of which SCC comprised only 52%. Malignant lymphoma most frequently occurred in the oropharynx, while sarcomas and melanomas preferred the oral cavity and sinonasal tract. Relative incidence of SCC among head and neck cancers was lower, and those of sarcoma and malignant melanoma were higher in Korea than in the North America. The etiologic and epidemiologic study on sarcomas and melanomas of the head and neck is required.

The Risk of Upper Aero Digestive Tract Cancer associated with Smoking, with and without Concurrent Alcohol Consumption

Smoking and alcohol are major causal factors for upper aerodigestive tract cancer, but reliable quantification of the combined impact of smoking and alcohol on this cancer and its major subtypes has not been performed. A meta-analysis of studies that had published quantitative estimates of smoking and upper aerodigestive tract cancer by January 2007 was performed. Pooled estimates of relative risks were obtained. Publication bias was investigated through funnel plots and corrected if found to be present. Overall, 85 studies with information on 53,940 individuals with upper aerodigestive tract cancer were included. The pooled estimate for the association between smoking and the risk of this cancer was 3.47 (95% confidence interval, 3.06-3.92). The risk remained elevated for a decade after smoking cessation but declined thereafter. Individuals who both smoked and consumed alcohol had double the risk of upper aerodigestive tract cancer in comparison with those who only smoked: the relative risk was 6.93 (95% confidence interval, 4.99-9.62) for the former and 2.56 (95% confidence interval, 2.20-2.97) for the latter (P < 0.001). Public health interventions that simultaneously discourage smoking and heavy drinking would have greater benefits than would be expected from those that target only one of these risk factors.

Quantitative expression and immunogenicity of MAGE‐3 and ‐6 in upper aerodigestive tract cancer

The MAGE antigens are frequently expressed cancer vaccine targets. However, quantitative analysis of MAGE expression in upper aerodigestive tract (UADT) tumor cells and its association with T-cell recognition has not been performed, hindering the selection of appropriate candidates for MAGE-specific immunotherapy. Using quantitative RT-PCR (QRT-PCR), we evaluated the expression of MAGE-3/6 in 65 UADT cancers, 48 normal samples from tumor matched sites and 7 HLA-A*0201+ squamous cell carcinoma of the head and neck (SCCHN) cell lines. Expression results were confirmed using Western blot. HLA-A*0201:MAGE-3- (271-279) specific cytotoxic T lymphocytes (MAGE-CTL) from SCCHN patients and healthy donors showed that MAGE-3/6 expression was highly associated with CTL recognition in vitro. On the basis of the MAGE-3/6 expression, we could identify 31 (47%) of the 65 UADT tumors, which appeared to express MAGE-3/6 at levels that correlated with efficient CTL recognition. To confirm that the level of MAGE-3 expression was responsible for CTL recognition, 2 MAGE-3/6 mRNA(high) SCCHN cell lines, PCI-13 and PCI-30, were subjected to MAGE-3/6-specific knockdown. RNAi-transfected cells showed that MAGE expression and MAGE-CTL recognition were significantly reduced. Furthermore, treatment of cells expressing low MAGE-3/6 mRNA with a demethylating agent, 5-aza-2'-deoxycytidine (DAC), increased the expression of MAGE-3/6 and CTL recognition. Thus, using QRT-PCR UADT cancers frequently express MAGE-3/6 at levels sufficient for CTL recognition, supporting the use of a QRT-PCR-based assay for the selection of candidates likely to respond to MAGE-3/6 immunotherapy. Demethylating agents could increase the number of patients amenable for targeting epigenetically modified tumor antigens in vaccine trials.

Results of Esophagogastroduodenoscopy in Patients With Oral Squamous Cell Carcinoma—Value of Endoscopic Screening: 10-Year Experience

Oral squamous cell carcinoma (OSCC) represents more than 90% of oral malignancies. Risk factors such as smoking and alcohol abuse can result in additional diseases, particularly of the upper gastrointestinal tract. The aim of the present study was to assess the synchronous pathologic changes of the upper gastrointestinal tract using endoscopy at the diagnosis of OSCC using a retrospective cohort study design. The clinical records of 806 patients with OSCC from 1997 to 2007 were reviewed. Of the 806 patients, 570 (188 females and 382 males) with primary OSCC were identified. All patients underwent esophagogastroduodenoscopy at the initial tumor staging. The tumor stage was defined according to the International Union Against Cancer classification for head and neck malignancies. The clinical and histologic data were analyzed for probability using univariate and multivariate analyses. Of the 570 patients, 289 had limited and 281 advanced disease. A statistically significant association was found between the advanced and limited tumor stage for Barrett's esophagus with an odds ratio of 4.986 (95% confidence interval 1.370 to 27.360, P = .0061). Pathologic findings were observed in 169 patients. Additional risk factors for the development of secondary upper aerodigestive tract cancers were found in 103 patients. Esophagogastroduodenoscopy provides the opportunity to identify second malignancies, precancerous lesions, and risk factors and to obtain biopsies using one procedure. The results can be immediately integrated into the therapeutic concept of the primary disease. Although new imaging techniques have been available, esophagogastroduodenoscopy still plays a decisive role in the routine staging of OSCC.